Identification and Validation of References Genes for Transcriptome Analysis of Murine Gammaherpesvirus 68 Infection
Reddy, Sandeep Steven-Andrew
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The human gammaherpesviruses (gammaHV), Kaposi's sarcoma-associated virus (KSHV) and Epstein-Barr virus (EBV), are the etiologic agents of many lymphoproliferative and neoplastic disorders. Murine gammaherpesvirus 68 (MHV68) has a high biological and genetic correlation to EBV and KSHV and represents an amenable, tractable model system to investigate host-virus interactions during chronic gammaHV infections. Modulating host cellular gene expression is one common strategy utilized during infection by all three viruses. Here, we have shown that four previously reported housekeeping genes (HKGs), ActB, beta2M, GAPDH and HPRT, evade host shutoff and exhibit stable transcript expression. The validation of these genes in the context of MHV68 infection allowed us to normalize the transcriptional expression of viral ORF50, an immediate early gene, by RT-qPCR. The validation of these four genes will allow us to identify additional HKGs with similar attributes that escape virally induced host shutoff. The compilation of these validated and novel HKGs can then be applied as reference genes to define the transcriptome of gammaherpesvirus infections in multiple cell lines and phases of the viral lifecycle.